146-P

PLOS ONE, 2015

Over the recent years, next generation sequencing and microarray technologies have revolutionized scientific research with their applications to high-throughput analysis of biological systems. Isolation of high quantities of pure, intact, double stranded, highly concentrated, not contaminated genomic DNA is prerequisite for successful and reliable large scale genotyping analysis. High quantities of pure DNA are also required for the creation of DNA-banks. In the present study, eleven different DNA extraction procedures, including phenol-chloroform, silica and magnetic beads based extractions, were examined to ascertain their relative effectiveness for extracting DNA from ovine blood samples. The quality and quantity of the differentially extracted DNA was subsequently assessed by spectrophotometric measurements, Qubit measurements, real-time PCR amplifications and gel electrophoresis. Processing time, intensity of labor and cost for each method were also evaluated. Results revealed significant differences among the eleven procedures and only four of the methods yielded satisfactory outputs. These four methods, comprising three modified silica based commercial kits (Modified Blood, Modified Tissue, Modified Dx kits) and an in-house developed magnetic beads based protocol, were most appropriate for extracting high quality and quantity DNA suitable for large-scale microarray genotyping and also for long-term DNA storage as demonstrated by their successful application to 600 individuals.

Journal of Applied Oral Science, 2012

O bjective: The aim of this study was to evaluate, by PCR-RFLP and Real-time PCR, the yield and quality of genomic DNA collected from buccal cells by mouthwash after different storage times at room temperature. Material and Methods: A group of volunteers was recruited to collect buccal cells using a mouthwash solution. The collected solution was divided into 3 tubes, one tube were used for immediate extraction and the remaining received ethanol and were kept at room temperature for 4 and 8 days followed by DNA extraction. The concentration, purity and integrity of the DNA were determined using spectrophotometry and electrophoresis. DNA quality differences among the three incubation times were also evaluated for genotyping eGF +61 A/G (rs 4444903) polymorphism by PCR-RFLP and for IRF6 polymorphism (rs 17015215) using Real-Time PCR. Results: There was no significant difference of DNA yield (p=0.75) and purity (p=0.86) among the three different incubation times. DNA obtained from different incubation times presented highmolecular weight. The PCR-RFLP and Real time PCR reactions were successfully performed for all DNA samples, even those extracted after 8 days of incubation. All samples genotyped by Real-Time PCR presented C allele for IRF6 gene polymorphism (homozygous: CC; heterozygous: CT) and the C allele was used as a reference for Ct values. The samples presented the same genotype for the different times in both techniques. Conclusion: We demonstrated that the method described herein is simple and low cost, and that DNA can be extracted and PCR amplified after storage in mouthwash solution at room temperature.

2018

DNA extraction is one of the most modern of the biological sciences. Scientists and doctors use DNA extraction to diagnose many medical conditions. DNA extraction can also be used to gather evidence in a criminal investigation. There are many methods for DNA isolation from human samples but all the available methods are more time consuming and require corrosive chemicals like phenol-chloroform, liquid nitrogen, silica and they are highly expensive chemicals also or it requires high-cost kits. The present study describes a rapid, reliable SDS buffer protocol suitable for extraction of DNA from Blood, fresh human tissue, dried tissue, bone, paraffin-embedded tissue and plastinated tissue samples. The protocol does not any require toxic chemicals and also consumes less time (<1 hour) and also cost-effective. In our study extracted DNA showed the excellent purity evidenced by UV spectrometer at the ratio of A260/A280 ranging from 1.8-1.85 and A260/A230 ratio was 2, which shows the pr...

A large number of different protocols for the efficient isolation of highly purified DNA from eukaryotic and prokaryotic cells is extant. (1-4) These procedures usually include treatment with proteinase K in the presence of SDS, which efficiently lyses the cells and nuclei and liberates the DNA tightly bound in chromatin. (s) Proteins are then extracted with phenol and chloroform, and the nucleic acids are precipitated with ethanol. This procedure is tedious and time-consuming, and significant amounts of DNA may be lost, especially when working with small specimens (e.g., joint biopsies). Therefore, this approach is not appropriate for diagnostic tests. Direct amplification of digested samples without phenol/chloroform extraction and precipitation is not possible because SDS is inhibitory to Taq polymerase at concentrations as low as 0.01%. (6) Alternative simple DNA extraction procedures have been used but have often resulted in incomplete lysis of the cells. These procedures typically have included detergents (e.g., Triton X-100), chaotropes (e.g., guanidium isothiocyanate or sodium iodide), proteases (e.g., proteinase K), substances that lyse erythrocytes and leukocytes (e.g., saponin), or heat denaturation. Often nonionic detergents such as Tween 20 or Laureth 12 in combination with proteinase K are used, followed by heat inactivation of the enzyme prior to PCR amplification. (7-1°)

Open Veterinary Journal, 2018

DNA is the prerequisite for life's inception that transfers hereditary information, past several years; various types of commercial kits are made available which vary depending on the type of the biological sample being used. The present study is focused on developing an improvised methodology for the isolation of genomic DNA from stored bovine blood samples. DNA was isolated by using the conventional Phenol: Chloroform: Isoamyl alcohol (PCI) method and Detergent method. The aim of the study was to make a comparative analysis and evaluation of these two methods to identify the one that gives a superior quality and quantity of genomic DNA. Total (n=48) each duplicate blood samples from three different buffalo(Bubalus bubalis) breeds Banni, Surti, Murrah, three zebu cattle (Bos indicus) breeds Kankerj, Gir, Sahiwal were collected from the jugular vein. The quantity, purity of the genomic DNA was assessed based on the total DNA yield, purity ratios, spectral profile, agarose gel electrophoresis analysis and polymerase chain reaction amplification of MC1R gene product without any inhibitors. The results of our study suggest that detergent method is also suitable for extraction of genomic DNA from the bovine blood and results were significant (*P>0.05). The total mean yield was found to be 329.05±11 μg/5ml for all six breeds while the PCI method was employed. The total mean yield of the gDNA for all six breeds was 406.6±43 μg/5ml of blood when the detergent method was used. One way ANOVA test showed that the total DNA yield varied depending on the isolation method used. The DNA yield obtained from the DG method was (***P< 0.001) significant as compared to the PCI method (**P<0.01).

Forensic Science International: Genetics, 2014

Unité et diversité du monde celtique. - Unity and Diversity in the Celtic World Actes du 42e colloque international de l’AFEAF (Prague, 10-13 mai 2018), 2020

Significado del Ahorro de Energéticos en la Economía de las Industrias Mexicanas, 1989

Estrategias para el ahorro de energéticos y primeros pasos hacia la Sustentabilidad. Strategies for energy saving and first steps towards sustainability.

Le rogamos haga buen uso de este folleto cuando ya no le sea útil.

Invisibility in Visual and Material Culture, edited by Asbjørn Grønstad, Øyvind Vågnes. Palgrave Macmillan, 2019

The chapter focuses on the issue of transmedial and sensory exchange in the context of digital culture and biometric technology. It analyzes critically the epistemic claims behind the various brain-scanning technologies, focusing on the status of the inner images that underlie cognitive activity. Multimedia performances and artistic experiments designed in collaboration with neuroscientists open up new dimensions in the discussion of translation between different sensory modalities, as well as translation between human perceptive apparatus and computational systems. Engaging the methodologies of contemporary image science and critical neuroscience, the paper shows how artistic scenarios help to both localize and expand our understanding of mental imagery and to offer an alternative to the existing correlations-based approach.